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. 2024 Mar 1;300(4):107130. doi: 10.1016/j.jbc.2024.107130

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© 2024 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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p47^phoxbinding does not impact WRC activity.A, cogel filtration of WRC^Δ230 with glutathione-S-transferase (GST)-p47^phox AB, showing chromatograms of indicated samples on left and Coomassie blue–stained SDS-PAGE gels on right. Gel images are aligned vertically over indicated fractions. The formation of a WRC–p47^phox complex is indicated by the shift of the WRC peak from fraction 18 to 17 and the appearance of p47^phox in fractions 16 to 18. B, Coomassie blue–stained SDS-PAGE gels of amylose beads pull down using indicated MBP-tagged baits (41), comparing untagged versus GST-tagged p47^phox PAB^short. C–F, pyrene-actin polymerization assay at indicated conditions. Reactions contain 3 to 4 μM actin (5% pyrene-labeled), 10 nM Arp2/3, 0.1 μM WRC^230WCA or WCA, and indicated Rac1^Q61L and/or p47^phox. WCA, WH2-central-acidic; WRC, WAVE regulatory complex.